LOCUS Exported 9306 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FU-tet-o-hNanogP8 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9306) AUTHORS Maherali N, Ahfeldt T, Rigamonti A, Utikal J, Cowan C, Hochedlinger K TITLE A high-efficiency system for the generation and study of human induced pluripotent stem cells. JOURNAL Cell Stem Cell. 2008 Sep 11. 3(3):340-5. PUBMED 18786420 REFERENCE 2 (bases 1 to 9306) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9306 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2098..2142 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2610..2880 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 2913..2950 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2947..2971 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 3998..4586 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4051..4071) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4469..4480) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4589..4605) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4590..4606) /label=pBluescriptKS /note="For pBluescript vector" LTR 5111..5291 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5317..5334) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5323..5547 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5593..6021 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5680..5699) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5890..5911 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6030..6050) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6035..6364 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6215..6350 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6277..6296 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 6412..6459 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6478..6852 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 6982..7103 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7019..7038) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7073..7092 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7152..7168) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7152..7168) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7165..7187) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7176..7192 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(7200..7230) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7245..7266 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7383..7400) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7554..8142) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7634..7653) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8313..9173) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8936..8955 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9174..9278) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttatcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc 2641 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat 2701 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg 2761 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact 2821 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga 2881 aaagtgaaag tcgagctcgg tacccgggtc gagtaggcgt gtacggtggg aggcctatat 2941 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga 3001 cctccataga agacaccggg accgatccag cctccgcggc cccgaattcg ccaccatgag 3061 tgtggatcca gcttgtcccc aaagcttgcc ttgctttgaa gcatccgact gtaaagaatc 3121 ttcacctatg cctgtgattt gtgggcctga agaaaactat ccatccttgc aaatgtcttc 3181 tgctgagatg cctcacacag agactgtctc tcctcttcct tcctccatgg atctgcttat 3241 tcaggacagc cctgattctt ccaccagtcc caaaggcaaa caacccactt ctgcagagaa 3301 tagtgtcgca aaaaaggaag acaaggtccc ggtcaagaaa cagaagacca gaactgtgtt 3361 ctcttccacc cagctgtgtg tactcaatga tagatttcag agacagaaat acctcagcct 3421 ccagcagatg caagaactct ccaacatcct gaacctcagc tacaaacagg tgaagacctg 3481 gttccagaac cagagaatga aatctaagag gtggcagaaa aacaactggc cgaagaatag 3541 caatggtgtg acgcagaagg cctcagcacc tacctacccc agcctctact cttcctacca 3601 ccagggatgc ctggtgaacc cgactgggaa ccttccaatg tggagcaacc agacctggaa 3661 caattcaacc tggagcaacc agacccagaa catccagtcc tggagcaacc actcctggaa 3721 cactcagacc tggtgcaccc aatcctggaa caatcaggcc tggaacagtc ccttctataa 3781 ctgtggagag gaatctctgc agtcctgcat gcacttccag ccaaattctc ctgccagtga 3841 cttggaggct gccttggaag ctgctgggga aggccttaat gtaatacagc agaccactag 3901 gtattttagt actccacaaa ccatggattt attcctaaac tactccatga acatgcaacc 3961 tgaagacgtg tgagaattcg atatcaagct tatcgataat caacctctgg attacaaaat 4021 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 4081 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 4141 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 4201 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 4261 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 4321 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 4381 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 4441 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 4501 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 4561 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca 4621 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc 4681 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac 4741 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 4801 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta 4861 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg 4921 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga 4981 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt 5041 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc 5101 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 5161 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 5221 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 5281 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc 5341 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca 5401 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta 5461 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc 5521 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta 5581 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 5641 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 5701 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 5761 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 5821 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 5881 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 5941 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 6001 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc 6061 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 6121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 6181 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 6241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6301 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 6361 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat 6421 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg 6481 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag 6541 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg 6601 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac 6661 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc 6721 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg 6781 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag 6841 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg 6901 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 6961 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 7021 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 7081 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 7141 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 7201 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 7261 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 7321 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 7381 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 7441 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 7501 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 7561 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 7621 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 7681 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 7741 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 7801 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7861 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7921 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7981 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 8041 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 8101 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 8161 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 8221 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 8281 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 8341 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 8401 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 8461 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 8521 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 8581 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 8641 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 8701 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 8761 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 8821 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 8881 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 8941 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 9001 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 9061 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 9121 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 9181 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 9241 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 9301 cctgac //